KR20200078907A - Saccharomyces cerevisiae afy-5 strain with high productivity of flavor components and process for preparing alcoholic liquors using the same - Google Patents

Abstract

The present invention relates to a Saccharomyces cerevisiae AFY-5 yeast producing a fragrance component at high concentrations, and a manufacturing method of alcoholic beverages using the same. The Saccharomyces cerevisiae AFY-5 strain contains 26S rRNA consisting of the nucleotide sequence of SEQ ID NO: 1.

Description

Fragrance Ingredients High Productivity Saccharomyces cerevisiae AFY-5 Yeast and method of manufacturing alcoholic beverages using the same

Saccharomyces cerevisiae AFY-5, which produces fragrance components at high concentrations, and a method for producing alcoholic beverages using the same.

Saccharomyces cerevisiae strains are used in a variety of alcoholic beverages, including beer. Saccharomyces cerevisiae strains are known to exist in a variety of subtypes, and their properties are also different and diverse.

Many flavor components are involved in the taste and aroma of beer. Among them, urea and hydrogen sulfide, which can be produced by strains, can cause toxicity to mammals or reduce the flavor of beer. Other processes can be added or additives can be added in manufacturing to remove them, but they are not cost or time efficient. Also, the flavor of beer may be reduced by additional processes or additives.

A method of applying genetic modification may be used to obtain a beer yeast strain having excellent taste and aroma, but this requires a lot of time and labor, and even if the strain is selected in this way, the selected strains are often possessed by the original parent strain. There are many cases in which excellent fermentative properties have been lost, and there are disadvantages such as concerns about safety of genetically engineered yeast.

Therefore, it is necessary to obtain a safe strain while producing a high concentration of beneficial scent components in the production of alcoholic beverages, but not producing substances that cause unpleasant flavor.

One aspect of the present invention to provide a strain comprising a 26S rRNA consisting of the nucleotide sequence of SEQ ID NO: 1.

Another aspect of the present invention comprises the steps of inoculating the strain according to one aspect to the wort with absorbance OD ₆₀₀ value of 0.1 to 0.3; A first fermentation step of fermenting at 15°C to 45°C for 5 to 30 days; And a second fermentation step of adding sugars to ferment at 15°C to 45°C for 2 to 30 days.

Another aspect of the present invention is to provide a mainstream composition prepared using a strain according to one aspect.

All technical terms used in the present invention, unless defined otherwise, are used in the sense as commonly understood by those skilled in the art in the relevant field of the present invention. In addition, although a preferred method or sample is described herein, similar or equivalent ones are included in the scope of the present invention. In addition, the numerical values described in this specification are considered to include the meaning of “about” even if not specified. The contents of all publications described by reference herein are incorporated herein by reference in their entirety.

One aspect provides a strain comprising 26S rRNA consisting of the nucleotide sequence of SEQ ID NO: 1.

The strain may be yeast, Saccharomyces cerevisiae , and more specifically, Saccharomyces cerevisiae AFY-5. Specifically, the 26S rRNA may be an rRNA transcribed from a gene represented by the nucleotide sequence of SEQ ID NO: 1.

In one embodiment, the strain may be deposited with accession number KACC 93320P.

In one embodiment, the strain may be for use in the production of alcoholic beverages. The liquor may be beer, Yakju, Cheongju, Takju, fruit wine, shochu, distilled spirit, or fermented alcohol, but is not limited thereto.

In one embodiment, the liquor may preferably be beer.

In one embodiment, the strain may be intended to be applied to culture conditions with a sugar content of 1 w/w% or more.

In one embodiment, the strain may be for applying to culture conditions of pH 3.0 or higher.

The culture conditions may refer to the fermentation conditions of the strain required for the production of alcoholic beverages. The sugar content is 1 w/w% or more, 5 w/w% or more, 10 w/w% or more, 15 w/w% or more, 20 w/w% or more, 30 w/w% or more, 40 w/w% or more Or more, 1 to 60 w/w%, 5 to 60 w/w%, 10 to 60 w/w%, 15 to 60 w/w%, 20 to 60 w/w%, 30 to 60 w/w%, 40 to 60 w/w%, 1 to 50 w/w%, 5 to 50 w/w%, 10 to 50 w/w%, 15 to 50 w/w%, 20 to 50 w/w%, 30 to 50 w/w%, 40 to 50 w/w%, or 1 to 40 w/w%. The sugar includes, but is not limited to, monosaccharides, disaccharides, oligosaccharides, polysaccharides, or derivatives thereof. The sugars include glucose, fructose, galactose, sucrose, lactose, maltose, trehalose, fructooligosaccharide, galactoligosaccharide, starch, glycogen, cellulose, chitin, pectin, stachyose, sorbose, xylose Os, ribose, myoinistos, or myoinisitol.

The strain according to one embodiment can be cultured even in the mainstream production conditions requiring a wide range of sugar content, including high concentration, and thus can be applied to various mainstream production methods. The high concentration is 80 w/w% or less, 70 w/w% or less, 60 w/w% or less, or 50 w/w% or less, while 20 w/w% or more, 30 w/w% or more, or 40 w/ It may mean a sugar concentration of w% or more. Acidity is pH 3.0 or higher, pH 3.5 or higher, pH 4.0 or higher, pH 4.5 or higher, pH 5.0 or higher, pH 5.5 or higher, pH 6.0 or higher, pH 3 to 8.0, pH 3.5 to 8.0, pH 4.0 to 8.0, pH 4.5 to 8.0, pH 5.0 to 8.0, pH 5.5 to 8.0, pH 6.0 to 8.0, pH 3.0 to 7.0, pH 3.5 to 7.0, pH 4.0 to 7.0, pH 4.5 to 7.0, pH 5.0 to 7.0, pH 5.5 to 7.0, pH 6.0 to 7.0, Or pH 3.0 to 6.0. The strain according to one embodiment can be cultured under a wide range of acidity conditions, so that it is easy to culture and is applicable to various alcoholic beverage manufacturing methods. In addition, the strain according to one embodiment can be cultured even at a low pH condition, thereby blocking the contaminant contamination, thereby obtaining stability of the fermentation strain. The low pH condition may mean pH 3.0 to 5.0, pH 3.0 to 4.0, or pH 3.0 to 3.5.

In one embodiment, the strain may be for application in the culture conditions of 15 ℃ to 45 ℃. The strain according to an embodiment has a wide growth temperature range from low temperature to high temperature, so that it is easy to culture and is applicable to various alcoholic beverage manufacturing methods. The low temperature may mean 15°C to 30°C, 15°C to 25°C, or 15°C to 20°C. The high temperature may mean 35°C to 45°C or 40°C to 45°C.

In one embodiment, the strain may be that does not produce urea (urea).

In one embodiment, the strain may be one having α-amylase (α-Amylase), β-glucosidase (β-glucosidase), protease, and glucanase activity.

The strain according to one embodiment does not produce urea, so the alcohol produced using the strain may not contain urea. The strain according to one embodiment has high activity of α-amylase, β-glucosidase, protease, and glucanase, and does not produce urea, a carcinogen, so both efficiency and stability are high.

In one embodiment, the strain may be one that does not produce hydrogen sulfide (H ₂ S). Therefore, the liquor produced by the strain may not contain hydrogen sulfide, and may not contain an unpleasant aroma derived from hydrogen sulfide during the fermentation process in the production of liquor.

In one embodiment, the strain may be resistant to cerulenin. Since the strain according to one embodiment is resistant to cerulenin, it may be to produce ethyl caproate at a high concentration. Ethyl caproate is a substance produced by the esterification reaction of capronic acid, which is a kind of fatty acid, and ethanol, which is the main component of alcohol, by brewing yeast. It is also known as an important ingredient in alcoholic beverages such as Takju. Ethyl caproate is a substance made of capronic acid and ethanol through an esterification reaction by yeast, and has a fruity scent.It is not only sake in Japan, but also in traditional alcoholic beverages such as Korean traditional distilled spirits, Takju, and Chinese white wine. It is a fragrance ingredient known as an important ingredient. Cerulenin, an antibiotic, specifically inhibits yeast fatty acid synthase (FAS) and consequently inhibits yeast growth. That is, it is known that yeast that is resistant to cerulenin does not inhibit FAS and thus proliferates in the medium to which cerulenin is added, and the content of ethyl caproate increases accordingly as the amount of heavy chain fatty acid increases (Eiji ICHIKAWA, 　et al. ,　Agric. Biol. Chem., 55(8), 2153-2154, 1991] and [ARIKAWA YUKIHIKO,　et　al.,　Journal　of　Bioscience　and　Bioengineering, 90(6), 675-677, 2000]). Therefore, high-quality liquor rich in flavor can be produced using the strain.

In one embodiment, the strain is one or more selected from the group consisting of glucose, galactose, alpha-methyl-D-glucoside (α-Methyl-D-Glucoside), D-maltose, D-saccharose and D-rapinose It may be using a carbon source. The strain according to one embodiment may use a carbon source as described above, but may not use D-trehalose. Since the strain has a different carbon source utilization ability than the general alcohol production strain, it has a differentiation in the flavor and quality of the alcohol produced by fermentation, and can be used to produce excellent alcohol.

In one embodiment, the strain is ethyl caproate, ethyl caprate, ethyl laurate, isoamyl decanoate, isoamyl caprylate ), decyl acetate, phenethyl acetate, and 2-phenylethanol may be to produce one or more components selected from the group consisting of high concentrations. The strain according to one embodiment may be capable of producing the compound at a concentration that is 1.5 times or more, 2 times or more, or 3 times or more higher than a conventionally known strain, for example, a WB-06 strain.

In one embodiment, one or more compounds selected from the group consisting of the ethyl caproate, ethyl caprate, ethyl laurate, isoamyl decanoate, decyl acetate, and phenethyl acetate may be produced two or more times, and ethyl capro Eight, ethyl laurate or combinations thereof can be produced three times or more.

Another aspect is a mainstream production strain comprising 26S rRNA consisting of the nucleotide sequence of SEQ ID NO: 1,

The strain comprises at least one carbon source selected from the group consisting of glucose, galactose, alpha-methyl-D-glucoside, D-maltose, D-saccharose and D-rapinose; Sugar content of 1 w/w% or more; PH pH 3.0 or higher; And it is for application to one or more culture conditions selected from the culture conditions of 15 ℃ to 45 ℃,

The strain does not contain urea and hydrogen sulfide produced from the strain, ethyl caproate, ethyl caprate, ethyl laurate, isoamyl decanoate, isoamyl caprylate, decyl acetate, phenethyl acetate, and 2 -It may be for preparing alcoholic beverages containing a high concentration of one or more compounds selected from the group consisting of phenylethanol.

Another aspect is the step of inoculating the strain according to one aspect in the wort with an absorbance (OD ₆₀₀ ) value of 0.1 to 0.3; A first fermentation step of fermenting at 15°C to 45°C for 5 to 30 days; And a second fermentation step of adding sugars to ferment at 15°C to 45°C for 2 to 30 days.

The absorbance (OD ₆₀₀ ) may be 0.1 to 0.3, 0.12 to 0.28, 0.15 to 0.25, or 0.18 to 0.22.

The term "wort" may mean a liquid saccharified by grinding malt and mixing with hot water.

The temperature of the first fermentation step is 15 ℃ to 45 ℃, 15 ℃ to 40 ℃, 15 ℃ to 35 ℃, 15 ℃ to 30 ℃, 15 ℃ to 25 ℃, 15 ℃ to 20 ℃, 15 ℃ to 19 ℃, 16 ℃ to 19 ℃, 17 ℃ to 19 ℃, or 17.5 ℃ to 18.5 ℃. The first fermentation step may be 5 to 30 days, 5 to 20 days, 5 to 15 days, 5 to 12 days, 5 to 10 days, or 6 to 8 days.

Sugars used in the second fermentation step include, but are not limited to, monosaccharides, disaccharides, oligosaccharides, polysaccharides, or derivatives thereof. The sugars include glucose, fructose, galactose, sucrose, lactose, maltose, trehalose, fructooligosaccharide, galactoligosaccharide, starch, glycogen, cellulose, chitin, pectin, stachyose, sorbose, xylose Os, ribose, myoinisitose, or myoinisitol. In one embodiment, the saccharide may be at least one selected from the group consisting of glucose, galactose, alpha-methyl-D-glucoside, D-maltose, D-sacrose and D-rapinose.

The temperature of the second fermentation step is 15 ℃ to 45 ℃, 15 ℃ to 40 ℃, 15 ℃ to 35 ℃, 15 ℃ to 30 ℃, 15 ℃ to 25 ℃, 15 ℃ to 23 ℃, 18 ℃ to 21 ℃, Or 19.5°C to 20.5°C. The second fermentation step may be 2 to 30 days, 2 to 20 days, 2 to 15 days, 2 to 12 days, 2 to 10 days, or 2 to 7 days.

In one embodiment, the method can be a method of making beer.

In one embodiment, the wort comprises: a first aging step of aging the malt at 43°C to 47°C for 5 to 15 minutes; A second aging step of aging at 50°C to 55°C for 30 minutes to 50 minutes; A third aging step of aging at 60°C to 65°C for 50 to 70 minutes; A fourth aging step of aging at 75°C to 80°C for 5 to 15 minutes; And adding hops to the aged product and heating for 50 to 70 minutes. The heating is performed at a temperature above the boiling point of the aged material in the aged material, for example, 80°C or higher, 100°C or higher, 80°C to 150°C, 80°C to 120°C, 100°C to 150°C, or 100°C to 150°C. It can mean adding temperature. The heating step may mean a boiling step. The method may further include filtering before adding hops to the aged product. The temperature, aging time or heating time at each step may vary depending on the production amount, malt condition, hop condition, and desired flavor.

Another aspect provides a mainstream composition prepared using a strain according to one aspect.

It is understood that any of the terms or elements mentioned for the above preparation method and mainstream composition, such as those mentioned in the description of the claimed strain, is as mentioned in the description of the previously claimed strain.

The strain according to one aspect can be cultured under a wide range of sugar, acidity, or temperature conditions, and thus can be used to prepare various alcoholic beverages. In addition, the strain can be used to produce safe and high-quality alcoholic beverages because it produces various fragrance components at high concentrations and does not produce harmful substances such as urea or hydrogen sulfide and causes of unpleasant fragrances.

By using the method for manufacturing alcoholic beverages according to another aspect, it is possible to produce alcoholic beverages of high quality, specifically beer.

1 shows a genetic diagram of a strain according to one embodiment as a phylogenetic diagram.

Hereinafter, the present invention will be described in more detail by examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereby.

Example One: Saccharomyces Ceremony AFY -5 Identification and property confirmation

1-1. Saccharomyces Ceremony AFY -5 sympathy

10 g of fruit, 10 g of yeast, and 10 mL of makgeolli each added to 90 mL of distilled water were added to 90 mL of distilled water and stirred for 1 hour to extract the ingredients of each raw material, and then inoculated with YPD broth. And incubated at 25°C for 2 hours. Thereafter, in order to suppress the growth of the fungus, after culturing at 25° C. for 48 hours in YPD agar medium to which 0.35% sodium propionate was added, a single colony was obtained, and each was further cultured. The DNA was extracted from the strain obtained from the culture of the single colony using the Yeast DNA prep kit, and the ITS site using LR0R (5'-ACCCGCTGAACTTAAGC -3') and LR7 (5'-TACTACCACCAAGATCT-3') primers. Was amplified by PCR. After purifying the amplified site, the base sequence was determined by requesting sequencing from Macrogen Co., Ltd. The AFY-5 strain is Saccharomyces cerevisiae ) strains appeared to be 99% consistent. The strain identified as above was named Saccharomyces cerevisiae AFY-5 and deposited it at the National Academy of Agricultural Science, a depository institution (Accession No.: KACC 93320P). The base sequence of the analyzed ITS site (26S rRNA base sequence) is shown in SEQ ID NO: 1.

1-2. Glucose tolerance , Acid resistance evaluation

To evaluate glucose tolerance, glucose and maltose were added in 1, 5, 10, 15, 20, 30, and 40 w/w%, respectively, in YP (yeast extract 1%, peptone 2%, agar 2%) medium, acid resistance YP (yeast extract 1%, peptone 2%, agar 2%) medium with pH adjusted to 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, and 6.0, respectively, was used to evaluate. All experiments were inoculated by punching the strain with a Repicator in a medium prepared in a 96-well plate, and after 48 hours of incubation at 25°C, the colony size (mm) was measured, and 1 mm was +, 2 mm was ++, 3 Table 1 below shows mm as +++, 4 mm as ++++, and 5 mm as +++++.

Glucose One 5 10 15 20 30 40 AFY-5 ++++ ++++ ++++ ++++ ++++ ++++ +++

maltose One 5 10 15 20 30 40 AFY-5 +++ ++++ ++++ +++ +++ +++ +++

As can be seen from Tables 1 and 2, AFY-5 strains were confirmed to have excellent brewability as sugar-resistant strains with excellent growth even in environments containing high concentrations of sugars such as 40% glucose and maltose, respectively.

pH 3.0 3.5 4.0 4.5 5.0 5.5 6.0 AFY-5 ++++ ++++ ++++ ++++ ++++ ++++ ++++

As can be seen from Table 3, AFY-5 strains were able to grow within a wide range of pH 3.0 to 6.0. In particular, it was confirmed that in the initial fermentation, strain growth is possible even at low pH conditions such as pH 3.0 to 5.0, and thus it is possible to secure the safety of the fermentation strain due to the favorable contamination of germs.

1-3. Evaluation of growth by temperature

For evaluation by temperature, after culturing for 48 hours at 15, 20, 25, 30, 35, 40, and 45°C in YP medium, colony size (mm) was measured.

℃ 15 20 25 30 35 40 45 AFY-5 ++++ ++++ ++++ ++++ ++++ ++++ +++

As can be seen from Table 4, it can be seen that the AFY-5 strain has a wide range of growth temperature from low temperature to high temperature, and thus has advantages in fermentation temperature manipulation.

1-4. Enzyme activity evaluation

Enzyme activity for four enzymes, α-amylase (α-Amylase), β-glucosidase, protease, and glucanase, was performed by agar plate method. α-amylase enzyme activity was not stained by staining with iodine solution consisting of 1% KI and 0.1% KI ₂ after punching and inoculating the strain in YP agar with 1% (w/v) soluble starch added. Colonies were determined as α-amylase enzyme-producing strains. For β-glucosidase activity, 0.5% esculin was added, followed by sterilization of YPD agar adjusted to pH 5.0, followed by addition of 2% of 1% ferric ammonium citrate solution filtered to 0.45 μm. Medium was prepared. The strain was inoculated into the medium prepared as described above. The strain having a black ring around the colony was judged as a β-glucosidase positive strain. The protease enzyme activity was determined as a protease-positive strain by inoculating the strain in YM agar with 1% skim milk added to form a transparent ring around the colony. In the glucanase experiment, a strain inoculated with YPD agar medium in which 0.2% β-glucan was added was inoculated and cultured, and then stained with 0.1% congo red reagent to determine a strain in which a clear ring around the colony was produced was a glucanase positive strain. . Urease (Urease) was inoculated into Christensen's urea agar, and a strain in which a pink ring was generated around the colony was determined as a urease-positive strain.

Enzyme activity α-amylase β-glucosidase Protease Glucanase Eurease AFY-5 +++ +++++ +++ ++ -

As shown in Table 5, AFY-5 strain has high starch degrading enzyme activity such as α-amylase, β-glucosidase, protease, and glucanase, and fermentation strains can be prepared using various starch and protein raw materials. , It can be seen that as a urea-negative strain, urea, which is a carcinogen, is not produced.

1-5. H ₂ S Create and Cerulenin ( cerulenin ) Immunity evaluation

In order to check whether hydrogen sulfide was produced, the strain was inoculated into the Biggy agar and cultured at 25°C for 48 hours, and then the colony color was confirmed to be positive for the dark brown color and negative for the white color. Cerulenin resistance was evaluated in order to select an ethyl caproate high-producing strain. After adjusting the pH to 5.3 by adding 2% glucose and 2% agar to a 0.37% yeast nitrogen base in the fatty acid synthesis path, the size of the colonies was measured by inoculating the medium with 25 μm cerulenin.

division H ₂ S Cerulenin AFY-5 - +++++

As can be seen from Table 6, the AFY-5 strain was confirmed to be a non-hydrogen sulfide gas-producing strain and does not produce hydrogen sulfide, which produces an unpleasant aroma of fermentation wine, and is found to be resistant to cerulenin, thereby producing large amounts of ethyl caproate. It is judged that it will be done, and it can be seen that it is effective in manufacturing high-quality alcohol rich in flavor.

Example 2: Saccharomyces Ceremony AFY -5 Check the biochemical properties of the strain

The identified Saccharomyces cerevisiae AFY-5 strain was evaluated for carbon source availability using the API 20C AUX kit, and the results are shown in Table 7 below. Saccharomyces cerevisiae WB-06 (Fermentis Co.) was used as a comparative strain.

division WB-06 AFY-5 Control - - Glucose + + Glycerol - - 2-Keto-D-glucose - - L-Arabinose - - D-Xylose - - Adonitol - - Xylitol - - D-Galactose + + Inositol - - D-Sorbitol - - α-Methyl-D-Glucoside - + N-Acetyl-D-Glucosamine - - D-Celiobiose - - D-Lactose - - D-Maltose + + D-Saccharose + + D-Trehalose + - D-Melezitose - - D-Raffinose - + Hyphae/Pseudo-Hyphae - -

As can be seen from Table 7, WB-06 and AFY-5 strains are both Saccharomyces cerevisiae strain strains, but the carbon source availability is different. The ability to utilize these different carbon sources will help fermentation products to be obtained differently.

Example 3. Saccharomyces Ceremony AFY Beer fermentation using -5 strain

3-1. Preparation of wort

Malt (Pilsner, wheat, Charahell, acidulated malt) is added to the saccharification tank for about 10 minutes at about 45℃, about 40 minutes at about 52℃, about 60 minutes at about 63℃, about 10 at about 78℃ Minutes were treated. After the primary and secondary filtration, hops (Hallertaur, Cascade, Saaz) were added to the boiling vessel, followed by boiling for 60 minutes.

3-2. Beer manufacturing

WB-06 and AFY-5 strains were inoculated with wort so that the absorbance (OD ₆₀₀ ) was about 0.2, and fermentation was performed at about 18°C for about 7 days. The produced young beer was transferred to a 500 mL pressure resistant PET bottle for carbonic acid production, sugar was added, and fermented for 2 to 7 days at about 20°C for 2 to 7 days, and the finished beer was stored at 0°C.

3-3. Fragrance analysis

The aroma component is 5 mL of the beer prepared in Example 3-2 in 10 mL vials, equilibrated for 10 minutes at 50° C., collected on 50/30 μm DVB/CAR/PDMS fibers, and SPME (Solid Phase Microextraction). ) To GC-TOFMS (Leco, Pegasus HT, USA). The column was used Rxi-5ms (30 m × 0.25 mm, 0.25 μm), the sample was incubated at 50° C. for 10 minutes, and then extracted at 50° C. for 10 minutes. As a carrier gas, the flow rate was set to 1.0 mL/min using He gas, the oven temperature was maintained at 40°C for 2 minutes, and then heated to 250°C at a rate of 10°C/min, and then maintained for 2 minutes. The transfer line temperature was 250°C. The mass spectrum range was analyzed as 33 to 510 m/z, and the area% of the main fragrance components was calculated and shown in Table 8 below.

compound Fragrance description WB-06 strain AFY-5 strain Ethyl caproate Sweet, fruity, pineapple, waxy, green, banana, fat 0.136 0.434 Ethyl caprate Sweetness, waxy, fruit, apple, grape, oily, brandy 0.071 0.162 Ethyl laurate Wax, creamy, soap and flower flavors, dairy and fruit flavors 0.131 0.339 Isoamyl Decanoate Wax, fruit, grass, banana, creamy, cheese, fat 0.008 0.023 Isoamyl caprylate Sweetness, oily, fruit, herb, soap, pineapple, coconut 0.042 0.067 Decyl acetate Tangerine, apple, melon, strawberry, celery, and other edible substances, decyl acetate is apple, orange and rum flavor 0.007 0.015 Phenethyl acetate With a slightly juicy scent, flowers, roses, sweetness, honey, fruit, tropical fruit, and flavor 0.115 0.238 2-phenylethanol Flowers, dried equipment incense, rose water 0.131 0.243

As can be seen from Table 8, when preparing a beer using the AFY-5 strain, it was confirmed that most of the analyzed fragrance components contained 0.5 to 3 times or more compared to the case of using the control WB-06 strain Did.

So far, the present invention has been focused on preferred embodiments. Those skilled in the art to which the present invention pertains will understand that the present invention may be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an explanatory point of view rather than a restrictive point of view. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent range should be interpreted as being included in the present invention.

Depository name: Institute for Agricultural Biotechnology

Accession number: KACC93320P

Date of accession: 20181214

<110> KANGWON PROVINCE <120> SACCHAROMYCES CEREVISIAE AFY-5 STRAIN WITH HIGH PRODUCTIVITY OF FLAVOR COMPONENTS AND PROCESS FOR PREPARING ALCOHOLIC LIQUORS USING THE SAME <130> PAA-18-0108-KR0 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 849 <212> DNA <213> Saccharomyces cerevisiae <400> 1 taaagctaaa tattggcgag agaccgatag cgaacaagta cagtgatgga aagatgaaaa 60 gaactttgaa aagagagtga aaaagtacgt gaaattgttg aaagggaagg gcatttgatc 120 agacatggtg ttttgtgccc tctgctcctt gtgggtaggg gaatctcgca tttcactggg 180 ccagcatcag ttttggtggc aggataaatc cataggaatg tagcttgcct cggtaagtat 240 tatagcctgt gggaatactg ccagctggga ctgaggactg cgacgtaagt caaggatgct 300 ggcataatgg ttatatgccg cccgtcttga aacacggacc aaggagtcta acgtctatgc 360 gagtgtttgg gtgtaaaacc catacgcgta atgaaagtga acgtaggttg gggcctcgca 420 agaggtgcac aatcgaccga tcctgatgtc ttcggatgga tttgagtaag agcatagctg 480 ttgggacccg aaagatggtg aactatgcct gaatagggtg aagccagagg aaactctggt 540 ggaggctcgt agcggttctg acgtgcaaat cgatcgtcga atttgggtat aggggcgaaa 600 gactaatcga accatctagt agctggttcc tgccgaagtt tccctcagga tagcagaagc 660 tcgtatcagt tttatgaggt aaagcgaatg attagaggtt ccggggtcga aatgaccttg 720 acctattctc aaactttaaa tatgtaagaa gtccttgtta cttaattgaa cgtggacatt 780 tgaatgaaga gcttttagtg ggccattttt ggtaagcaga actggcgatg cgggatgaac 840 cgaacgtag 849

Claims (18)

Saccharomyces cerevisiae AFY-5 strain comprising 26S rRNA consisting of the nucleotide sequence of SEQ ID NO: 1. The method according to claim 1, The strain is deposited with the accession number KACC 93320P strain. The method according to claim 1, The strain is a strain for liquor production. The strain according to claim 1, wherein the alcoholic beverage is beer. The method according to claim 1, The strain is intended to be applied to culture conditions of sugar 1 w / w% or more. The method according to claim 1, wherein the strain is intended to be applied to culture conditions of pH pH 3.0 or higher. The method according to claim 1, The strain is intended to be applied in the culture conditions of 15 ℃ to 45 ℃. The method according to claim 1, wherein the strain is a strain that does not produce urea (urea). The strain according to claim 1, wherein the strain has α-amylase (α-Amylase), β-glucosidase (β-glucosidase), protease, and glucanase activity. The method according to claim 1, wherein the strain does not produce hydrogen sulfide (H ₂ S). The method according to claim 1, The strain is a strain that is resistant to cerulenin (cerulenin). The method according to claim 1, The strain is at least one selected from the group consisting of glucose, galactose, alpha-methyl-D-glucoside (α-Methyl-D-Glucoside), D-maltose, D-saccharose and D-rapinose A strain that uses a carbon source. The method according to claim 1, wherein the strain is ethyl caproate (ethyl caproate), ethyl caprate (ethyl caprate), ethyl laurate (ethyl laurate), isoamyl decanoate (isoamyl decanoate), isoamyl caprylate (isoamyl caprylate) ), decyl acetate, phenethyl acetate, and 2-phenylethanol may be to produce one or more components selected from the group consisting of high concentrations. A strain for mainstream production comprising 26S rRNA consisting of the nucleotide sequence of SEQ ID NO: 1,
The strain may include one or more carbon sources selected from the group consisting of glucose, galactose, alpha-methyl-D-glucoside, D-maltose, D-sacrose and D-rapinose; Sugar content of 1 w/w% or more; PH pH 3.0 or higher; And it is for application to one or more culture conditions selected from the culture conditions of 15 ℃ to 45 ℃,
The strain does not contain urea and hydrogen sulfide produced from the strain, ethyl caproate, ethyl caprate, ethyl laurate, isoamyl decanoate, isoamyl caprylate, decyl acetate, phenethyl acetate, and 2 -A strain for preparing alcoholic beverages comprising at least one compound selected from the group consisting of phenylethanol. Inoculating the strain of claim 1 with wort at an absorbance (OD ₆₀₀ ) value of 0.1 to 0.3;
A first fermentation step of fermenting at 15°C to 45°C for 5 to 30 days; And
Liquor production method comprising a second fermentation step of fermentation for 2 to 30 days at 15 ℃ to 45 ℃ by adding sugars. 16. The method of claim 15, wherein the liquor is beer. The method according to claim 15, The wort, A first aging step of aging the malt for 5 to 15 minutes at 43 ℃ to 47 ℃;
A second aging step of aging at 50°C to 55°C for 30 minutes to 50 minutes;
A third aging step of aging at 60°C to 65°C for 50 to 70 minutes;
A fourth aging step of aging at 75°C to 80°C for 5 to 15 minutes; And
The step of adding hops to the aged product and heating for 50 to 70 minutes may be further included. Liquor composition prepared using the strain of claim 1.

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